Successive capture of nucleic acid by magnetic glass particles

Abstract

Provided herein are methods and components for successive capture of nucleic acids using magnetic glass particles.

What is claimed: 
     
       1. A method for capturing nucleic acids in a liquid sample comprising:
 (a) contacting a first aliquot of the liquid sample with magnetic glass particles (MGPs) in a vessel under conditions that allow nucleic acids from the liquid sample to non-covalently bind to MGPs, wherein the MGPs are non-porous and comprise at least one ferromagnetic magnetic core in glass; 
 (b) applying a magnetic field to the MGPs; 
 (c) removing unbound liquid sample from the MGPs; 
 (d) contacting a second aliquot of the liquid sample with the MGPs; 
 (e) resuspending the MGPs in the second aliquot of the liquid sample; 
 (f) pipetting the MGPs to the top of the second aliquot and dispensing the MGPs; 
 (g) applying a magnetic field to the MGPs to form a clump of MGPs; 
 (h) removing unbound liquid sample from the clump of MGPs; and 
 (i) optionally repeating steps (d) through (h) for at least one additional aliquot of liquid sample. 
 
     
     
       2. The method of  claim 1 , wherein the MGPs have a mean diameter between 0.5-15 μm. 
     
     
       3. The method of  claim 1 , wherein the glass comprises at least one metal oxide. 
     
     
       4. The method of  claim 3 , wherein the at least one metal oxide is selected from SiO 2 , B 2 O 3 , Al 2 O 3 , K 2 O, CaO, and ZnO. 
     
     
       5. The method of  claim 3 , wherein the liquid sample is blood, plasma, serum, urine, or a lysate thereof. 
     
     
       6. The method of  claim 3 , wherein the liquid sample has a volume of at least 2 ml. 
     
     
       7. The method of  claim 6 , wherein the liquid sample has a volume of 2 ml to 100 ml. 
     
     
       8. The method of  claim 6 , wherein the vessel holds a volume of 2 ml or less. 
     
     
       9. The method of  claim 6 , wherein the vessel holds a volume of 0.5 ml to 2 ml. 
     
     
       10. The method of  claim 6 , further comprising eluting and separating the nucleic acids from the MGPs. 
     
     
       11. The method of  claim 6 , wherein the method is automated. 
     
     
       12. The method of  claim 6 , wherein step (e) comprises pipetting the MGPs in the liquid sample. 
     
     
       13. The method of  claim 6 , wherein the nucleic acids are RNA. 
     
     
       14. The method of  claim 6 , wherein the nucleic acids are DNA. 
     
     
       15. The method of  claim 6 , wherein the vessel is a well in a multi-well plate or a tube. 
     
     
       16. The method of  claim 6 , wherein step (c) comprises removing unbound liquid sample from the MGPs, washing the MGPs, and removing unbound material from the MGPs. 
     
     
       17. The method of  claim 6 , wherein step (h) comprises removing unbound liquid sample from the MGPs, washing the MGPs, and removing unbound material from the MGPs.